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Specificity and sensitivity of the LAMP-CRISPR/Cas12a assay

XW Xiang Wang
LW Lai-Fa Wang
YC Ye-Fan Cao
YY Yan-Zhi Yuan
JH Jian Hu
ZC Zu-Hai Chen
FZ Fei Zhu
XW Xi-Zhuo Wang
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The analytical specificity of the LAMP-CRISPR/Cas12a assay was tested on at least 5 ng of templates extracted from four B. xylophilus strains and three related species, B. mucronatus, B. doui, and Botrytis cinerea.

The analytical sensitivity for detecting a decreasing number of gene copies was evaluated using purified B. xylophilus DNA concentrations diluted from 10−1 to 10−8 to give DNA concentrations of 66.4 ng/μL, 6.64 ng/μL, 664 pg/μL, 66.4 pg/μL, 6.64 pg/μL, 0.664 pg/μL, 66.4 fg/μL, and 6.64 fg/μL.

We perform base alterations or deletions at one or more locations of the target DNA sequence to test that CRISPR-Cas12a enhanced fluorescence analysis can distinguish several base changes ( Supplementary Table S1 ).

Copyright and License information:  ©2022 Wang, Wang, Cao, Yuan, Hu, Chen, Zhu and Wang
This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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